Specific release of the extrinsic 18-kDa protein from spinach Photosystem-II particles by the treatment with NaCl and methanol and its application for large-scale purification of the three extrinsic proteins of Photosystem II without chromatography |
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| Affiliation: | 1. Shenzhen Engineering Laboratory for Marine Algal Biotechnology, Guangdong Technology Research Center for Marine Algal Biotechnology, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, China;2. College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China;3. Shenzhen Collaborative Innovation Public Service Platform for Marine Algae Industry, Longhua Innovation Institute for Biotechnology, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, China;4. The UWA Institute of Agriculture, The University of Western Australia, Crawley, Perth 6009, Australia |
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| Abstract: | The extrinsic 18-kDa protein in spinach Photosystem-II particles was specifically released from the membrane by the treatment with 0.5 M NaCl and 20% methanol at pH 6.5. The NaCl-methanol treatment was used in combination with the treatments with 2 M NaCl (pH 6.5) and 0.8 M Tris-HCl (pH 8.4) for developing a new procedure for the purification of the subunit proteins (the extrinsic 33-, 24- and 18-kDa proteins) of the oxygen-evolution enzyme complex from spinach chloroplasts. The three extrinsic proteins were liberated from the membranes almost completely and specifically by the simple washing procedure employed here. As no chromatographic step was required for the purification of the proteins, the time for the purification was considerably shortened and the yields of the proteins, especially of the 24- and 18-kDa proteins, were significantly improved. |
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